Transplantation of bone marrow mesenchymal stem cells overexpressing Shootin1 for treatment of spinal cord injury / 中国组织工程研究

ABSTRACT

BACKGROUND:Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cel s (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cel transplantation. OBJECTIVE:To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS:BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cel s under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Al en’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the model group were given no treatment. Five weeks after modeling, spinal cord samples were taken from each rat to make frozen sections for detection of nerve related markers RESULTS AND CONCLUSION:After 48-hour adenoviral transfection, Shootin1 expression was successful y detected in BMSCs. After 7-day in vitro induction, the cel morphology in the three groups varied, and there was no significant difference in the expression of Nestin and NeuN between the transfection and non-transfection groups. Basso, Beattie and Bresnahan scores were higher in the two cel transplantation groups than the model group. Increased expression levels of Nestin, NeuN, GFAP, MAP-2, ChAT and SYN were observed in both two cel transplantation groups, indicating a strengthened ability of nerve regeneration. Our experimental findings further confirm that BMSCs transplantation for spinal cord injury has achieved good outcomes, and Shootin1 protein plays a certain role in nerve regeneration and functional recovery after spinal cord injury. However, Shootin1 overexpression shows no obvious additional effects in combination with BMSCs transplantation, and further studies on the optimization of BMSCs transplantation for spinal cord injury are necessary.